Human Goodpasture anti-alpha3(IV)NC1 autoantibodies share structural determinants.
To examine the structural relationship among autoantibodies produced by individuals with anti-GBM antibody-mediated disease, a polyclonal anti-idiotype directed against human anti-alpha3(IV)NC1 antibodies was produced and then used to study autoantibodies from other patients. For this purpose, anti-alpha3(IV)NC1 antibodies (anti-GBM), derived from a single patient (LL) with high titer and typical anti-GBM antibody specificity, were isolated using recombinant alpha3(IV)NC1-sepharose affinity chromatography. Following hyperimmunization of rabbits with anti-GBM IgG, irrelevant rabbit anti-human IgG antibodies were removed from the antiserum using a human IgG-sepharose column. The rabbit anti-alpha3(IV)NC1 antibodies (anti-Id GBM) effluent bound to human anti-GBM antibodies, but it did not bind to either normal human IgG or recombinant alpha3(IV)NC1 protein. The Id-anti-Id interaction was conformationally dependent on intact heavy and light chains of the anti-alpha3(IV)NC1 antibodies (ELISA and Western blotting). A competitive immunoassay was developed to evaluate structural and potential genetic relationships among anti-alpha3(IV)NC1 antibodies from different patients. All patients tested (9 of 9) had a substantial fraction (producing > 50% inhibition) of anti-GBM antibodies expressing Id-GBM. The results indicate that shared determinants are expressed by anti-GBM antibodies from different individuals, and they raise the possibility that common genetic elements are used to encode them. These regions are potential targets for design of reagents to regulate autoreactive B cells and/or interfere with pathogenic antibody-GBM interactions, in individuals with anti-GBM antibody mediated diseases.
Meyers
K E
KE
Penn Center for Molecular Study of Kidney Diseases, The Children's Hospital of Philadelphia, University of Pennsylvania, 19104-6144, USA.
Kinniry
P A
PA
Kalluri
R
R
Neilson
E G
EG
Madaio
M P
MP
D000818:Animals; D019867:Anti-Glomerular Basement Membrane Disease / Q000276:immunology*; D000915:Antibody Affinity; D000937:Antigen-Antibody Reactions; D001323:Autoantibodies / Q000737:chemistry* / Q000378:metabolism; D001667:Binding, Competitive / Q000276:immunology; D003094:Collagen / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D015870:Gene Expression / Q000276:immunology; D006801:Humans; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D001323:Autoantibodies; D000939:Epitopes; D011994:Recombinant Proteins; D003094:Collagen